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Validation of Internal Control Genes for Quantitative Real-Time PCR Gene Expression Analysis in Morchella

Summary

This research focused on improving the accuracy of genetic studies in morel mushrooms by identifying reliable reference genes for measuring gene expression. This is important because morels are valuable edible mushrooms with medicinal properties, but studying their genetics has been challenging. The study’s impact on everyday life includes: • Better understanding of morel mushroom genetics could lead to improved cultivation methods • More reliable genetic research tools could help develop enhanced morel varieties • Improved knowledge may lead to better extraction of beneficial compounds from morels • Could help make morel mushrooms more widely available and affordable • Advances the potential for developing new medicines from morel compounds

Background

True morels (Morchella spp.) are edible mushrooms appreciated worldwide for their savory flavor and bioactive properties including anti-oxidative, anti-inflammatory, antimicrobial, immunostimulatory and antitumor effects. While cultivation technology has advanced, there remains limited molecular biological research on morels, partly due to challenges in completing their life cycle under laboratory conditions. Quantitative real-time PCR (qRT-PCR) requires reliable reference genes for normalization, but no universal reference gene maintains constant expression across all experimental conditions.

Objective

This study aimed to identify and validate stable reference genes for qRT-PCR normalization in Morchella under different developmental stages, carbon sources, and temperature stress conditions. The researchers sought to evaluate candidate genes across multiple Morchella species to establish reliable internal controls for gene expression analysis.

Results

ACT1 and INTF7 were identified as the most stable reference genes across all conditions. For specific conditions: CYC3 was most stable across developmental stages, INTF4/AEF3 performed best for carbon source variations, and INTF3/CYC3 showed robust stability under temperature stress. The study validated that ACT1, AEF3, CYC3, INTF3, INTF4 and INTF7 could serve as reliable reference genes across the 10 Morchella strains tested. Traditional reference genes like GPD2, BTU1 and ATU were found unsuitable as internal controls.

Conclusion

The study successfully identified and validated several novel reference genes for qRT-PCR analysis in Morchella species. The stability of reference genes varied under different experimental conditions, highlighting the importance of selecting specific reference genes for particular experimental conditions. This work provides a foundation for accurate gene expression analysis in Morchella and contributes to advancing molecular research in this important edible mushroom genus.
  • Published in:Molecules,
  • Study Type:Laboratory Research,
  • Source: 10.3390/molecules23092331
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